Prostatic cancer diagnostic kit and preparation method and application thereof

A diagnostic kit and prostate cancer technology, applied in the field of kits, can solve problems such as misdiagnosis and missed diagnosis of prostate cancer, and achieve high sensitivity and early diagnosis accuracy

Active Publication Date: 2019-05-17
杭州京北生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to provide a prostate cancer diagnostic kit that improves the diagnostic accuracy of prostate cancer, is fast, simple and highly sensitive in order to solve the defects that the existing detection of prostate cancer is easy to cause misdiagnosis and missed diagnosis.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0047] A diagnostic kit for prostate cancer, the kit contains:

[0048] 1. Luciferase-NPM1 antigen fusion protein;

[0049] 2. Buffer I;

[0050] 3. NPM1 antigen-coated plate;

[0051] 4. Renilla luciferase substrate;

[0052] 5. Substrate buffer;

[0053] 6. NPM1 antibody positive control substance;

[0054] 7. NPM1 antibody negative control substance.

[0055] The preparation method of the prostate cancer diagnostic kit is characterized in that the preparation method comprises the following steps: preparing luciferase-antibody fusion protein

[0056] Recombinant plasmid construction: According to the NPM1 antigen sequence, primers were designed and synthesized,

[0057] Upstream primer 5′CGGGGTACCCCG ATGGAAGATTCGATGGACATGGACA 3′,

[0058] Downstream primer 5′ TGCTCTAGAGCA TCAATGCGCTTTTTCTATACTTGCT 3′;

[0059] The anti-NPM1 gene was amplified using the serum of a patient with prostate cancer as a template. Kpnl and xbal were used to digest the multiple cloning site o...

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PUM

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Abstract

A kit is used for diagnosing NPM1 antibodies, ranilla luciferase genes and NPM1 antigen genes are recombined into fusion vectors, and through a cell transformation technology, the fusion vectors are guided into a mammalian cell system to conduct luciferase-NPM1 antigen fusion protein expression; gene recombination cells are disrupted, luciferase-NPM1 antigen fusion proteins are obtained, NPM1 antigens are coated with a 96 elisa plate, and to-be-detected serum is added; the antibodies in the serum are combined with the antigens on the plate to be adsorbed to the plate; and the luciferase-NPM1 antigen fusion proteins are added, through an immunoprecipitation reaction, the luciferase-NPM1 antigen fusion proteins are combined with the NPM1 antibodies in the serum of a patient and are adsorbedto the plate, and finally a luciferase substrate is added to detect the fluorescence intensity. According to the fluorescence intensity, the content of the NPM1 antibodies can be reflected to providereference for diagnosis of prostatic cancer; and a method is quick, simple and convenient and has high sensitivity.

Description

technical field [0001] The invention relates to a kit, in particular to a prostate cancer diagnostic kit which improves the diagnostic accuracy of prostate cancer, is fast, simple and high-sensitivity, and a preparation method and application thereof. Background technique [0002] Prostate cancer is a malignant tumor that occurs in the prostate tissue of men. It has become the second most common tumor in men worldwide and ranks sixth in cancer-related mortality. [0003] At present, the clinical diagnosis methods of prostate cancer mainly include prostate digital rectal examination (DRE), serum prostate-specific antigen (PSA) detection, rectal ultrasound detection, biopsy pathological examination and so on. [0004] Digital rectal examination is a classic technique for the diagnosis of prostate cancer. It mainly touches the prostate with the doctor's index finger to judge the size and shape of the prostate nodules, so as to determine whether there is prostate cancer. Biopsy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68G01N33/58G01N33/533
Inventor 唐东起周扬陈政鑫
Owner 杭州京北生物科技有限公司
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